{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T13:52:21Z","timestamp":1790085141003,"version":"4.0.1"},"reference-count":33,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,7,29]],"date-time":"2022-07-29T00:00:00Z","timestamp":1659052800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Key Research and Development Program of Hubei Province (CN)","award":["2020BBB079"],"award-info":[{"award-number":["2020BBB079"]}]},{"name":"Key Research and Development Program of Hubei Province (CN)","award":["2662022DKPY007"],"award-info":[{"award-number":["2662022DKPY007"]}]},{"name":"Key Research and Development Program of Hubei Province (CN)","award":["SZYJY2022024"],"award-info":[{"award-number":["SZYJY2022024"]}]},{"name":"Fundamental Research Funds for the Central Universities","award":["2020BBB079"],"award-info":[{"award-number":["2020BBB079"]}]},{"name":"Fundamental Research Funds for the Central Universities","award":["2662022DKPY007"],"award-info":[{"award-number":["2662022DKPY007"]}]},{"name":"Fundamental Research Funds for the Central Universities","award":["SZYJY2022024"],"award-info":[{"award-number":["SZYJY2022024"]}]},{"name":"HZAU-AGIS Cooperation Fund","award":["2020BBB079"],"award-info":[{"award-number":["2020BBB079"]}]},{"name":"HZAU-AGIS Cooperation Fund","award":["2662022DKPY007"],"award-info":[{"award-number":["2662022DKPY007"]}]},{"name":"HZAU-AGIS Cooperation Fund","award":["SZYJY2022024"],"award-info":[{"award-number":["SZYJY2022024"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>In the modern farming industry, the irrational or illegal use of veterinary drugs leads to residues in animal-derived food, which can seriously threaten human health. Efficient detection of low concentrations of drug residues in animal products in a short time is a key challenge for analytical methods. This study proposes to use an antibody chip biosensor for rapid and automated analysis of cephalosporins, aminoglycosides, and sulfonamide antibiotics in pork and milk. 3D polymer slides were applied for the preparation of antibody chips. Ovalbumin (OVA) or bovine serum albumin (BSA) conjugates of the haptens were immobilized as spots on disposable chips. Monoclonal antibodies (mAbs) against cefalexin, ceftiofur, gentamicin, neomycin, and sulfonamides allowed the simultaneous detection of the respective analytes. Antibody binding was detected by a second antibody labeled with Cy3-generating fluorescence, which was scanned a with chip scanner. The limits of detection (LOD) for all the analytes were far below the respective maximum residue limits (MRLs) and ranged from 0.51 to 4.3 \u00b5g\/kg. The average recoveries of all the analytes in each sample were in the range of 81.6\u2013113.6%. The intra- and inter-assay CV was less than 12.9% and showed good accuracy and precision for all the antibiotics at the MRL level. The sample pretreatment method is simple, and the results are confirmed to be accurate by LC\u2013MS\/MS; therefore, this method is valuable for the quality control of animal-derived food.<\/jats:p>","DOI":"10.3390\/bios12080578","type":"journal-article","created":{"date-parts":[[2022,7,31]],"date-time":"2022-07-31T21:49:02Z","timestamp":1659304142000},"page":"578","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["The Simultaneous Detection of Multiple Antibiotics in Milk and Pork Based on an Antibody Chip Biosensor"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9132-4294","authenticated-orcid":false,"given":"Jiaxu","family":"Xiao","sequence":"first","affiliation":[{"name":"National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nana","family":"Wei","sequence":"additional","affiliation":[{"name":"Kunpeng Institute of Modern Agriculture at Foshan, Foshan 528200, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuangmin","family":"Wu","sequence":"additional","affiliation":[{"name":"National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huaming","family":"Li","sequence":"additional","affiliation":[{"name":"National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyang","family":"Yin","sequence":"additional","affiliation":[{"name":"National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Si","sequence":"additional","affiliation":[{"name":"National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Long","family":"Li","sequence":"additional","affiliation":[{"name":"National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dapeng","family":"Peng","sequence":"additional","affiliation":[{"name":"National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"Shenzhen Institute of Nutrition and Health, Huazhong Agricultural University, Shenzhen 518000, China"},{"name":"Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China"},{"name":"Genome Analysis Laboratory of the Ministry of Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,29]]},"reference":[{"key":"ref_1","first-page":"1684","article-title":"Global Distribution of Extended Spectrum Cephalosporin and Carbapenem Resistance and Associated Resistance Markers in Escherichia coli of Swine Origin\u2014A Systematic Review and Meta-Analysis","volume":"13","author":"Hayer","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"126351","DOI":"10.1016\/j.chemosphere.2020.126351","article-title":"Occurrence and toxicity of antibiotics in the aquatic environment: A review","volume":"251","author":"Kovalakova","year":"2020","journal-title":"Chemosphere"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"7569","DOI":"10.1021\/acs.jafc.9b01334","article-title":"Antibiotic Residues in Food: Extraction, Analysis, and Human Health Concerns","volume":"67","author":"Chen","year":"2019","journal-title":"J. Agric. Food Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1093\/jac\/dkt443","article-title":"Correlation between veterinary antimicrobial use and antimicrobial resistance in food-producing animals: A report on seven countries","volume":"69","author":"Chantziaras","year":"2014","journal-title":"J. Antimicrob. Chemoth."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"906","DOI":"10.1016\/j.scitotenv.2017.09.328","article-title":"A duodecennial national synthesis of antibiotics in China\u2019s major rivers and seas (2005\u20132016)","volume":"615","author":"Li","year":"2018","journal-title":"Sci. Total Environ."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.chroma.2005.02.037","article-title":"Analytical strategies for residue analysis of veterinary drugs and growth-promoting agents in food-producing animals\u2014A review","volume":"1067","author":"Stolker","year":"2005","journal-title":"J. Chromatogr. A"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Rutjens, S., Croubels, S., De Baere, S., and Devreese, M. (2021). Development and Validation of Liquid Chromatography-Tandem Mass Spectrometry Methods for the Quantification of Cefquinome, Ceftiofur, and Desfuroylceftiofuracetamide in Porcine Feces with Emphasis on Analyte Stability. Molecules, 25.","DOI":"10.3390\/molecules26154598"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1016\/j.foodchem.2018.04.018","article-title":"Simultaneous determination of multi-residue and multi-class antibiotics in aquaculture shrimps by UPLC-MS\/MS","volume":"260","author":"Saxena","year":"2018","journal-title":"Food Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1510","DOI":"10.1080\/19440049.2017.1339331","article-title":"Strategies for the screening of antibiotic residues in eggs: Comparison of the validation of the classical microbiological method with an immunobiosensor method","volume":"34","author":"Gaudin","year":"2017","journal-title":"Food Addit. Contam. A"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1016\/j.idairyj.2013.04.004","article-title":"Microbiological system in microtitre plates for detection and classification of antibiotic residues in milk","volume":"32","author":"Nagel","year":"2013","journal-title":"Int. Dairy J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1870","DOI":"10.1080\/19440049.2013.832400","article-title":"Developing a microbiological growth inhibition screening assay for the detection of 27 veterinary drugs from 13 different classes in animal feedingstuffs","volume":"30","author":"Bohn","year":"2013","journal-title":"Food Addit. Contam. A"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.aca.2017.11.038","article-title":"Universal simultaneous multiplex ELISA of small molecules in milk based on dual luciferases","volume":"1001","author":"Yu","year":"2018","journal-title":"Anal. Chim. Acta"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"11170","DOI":"10.1021\/acs.jafc.8b03834","article-title":"Generic Hapten Synthesis, Broad-Specificity Monoclonal Antibodies Preparation, and Ultrasensitive ELISA for Five Antibacterial Synergists in Chicken and Milk","volume":"66","author":"Li","year":"2018","journal-title":"J. Agric. Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"830","DOI":"10.1080\/09540105.2016.1183599","article-title":"Establishment of a monoclonal antibody-based indirect enzyme-linked immunosorbent assay for the detection of trimethoprim residues in milk, honey, and fish samples","volume":"27","author":"Chen","year":"2016","journal-title":"Food Agric. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3163","DOI":"10.1007\/s12161-016-0508-4","article-title":"Microfluidic Chip-Based Immunoassay for Reliable Detection of Cloxacillin in Poultry","volume":"9","author":"Yu","year":"2016","journal-title":"Food Anal. Method"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1058","DOI":"10.1007\/s12161-014-9986-4","article-title":"Simultaneous Determination of Malachite Green, Diethylstilbestrol, Medroxyprogesterone, and 3-Amino-2-Oxazolidone in Synbranchoid Eels by a Protein Microarray Method","volume":"8","author":"Liu","year":"2015","journal-title":"Food Anal. Method"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1007\/978-1-4939-6584-7_13","article-title":"Validation Procedure for Multiplex Antibiotic Immunoassays Using Flow-Based Chemiluminescence Microarrays","volume":"1518","author":"Meyer","year":"2017","journal-title":"Methods Mol. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"4076","DOI":"10.1016\/j.bios.2011.03.036","article-title":"Simultaneous detection of four nitrofuran metabolites in honey using a multiplexing biochip screening assay","volume":"26","author":"Moloney","year":"2011","journal-title":"Biosens. Bioelectron."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.bios.2011.12.057","article-title":"Simultaneous and rapid detection of six different mycotoxins using an immunochip","volume":"34","author":"Wang","year":"2012","journal-title":"Biosens. Bioelectron."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"806","DOI":"10.1021\/acssensors.8b00051","article-title":"Disposable MoS2-Arrayed MALDI MS Chip for High-Throughput and Rapid Quantification of Sulfonamides in Multiple Real Samples","volume":"3","author":"Zhao","year":"2018","journal-title":"ACS Sens."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1016\/j.jchromb.2016.04.026","article-title":"Simultaneous determination of 22 cephalosporins drug residues in pork muscle using liquid chromatography-tandem mass spectrometry","volume":"1022","author":"Li","year":"2016","journal-title":"J. Chromatogr. B"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1016\/j.apsusc.2018.04.072","article-title":"Surface plasmon resonance imaging validation of small molecule drugs binding on target protein microarrays","volume":"450","author":"Zhou","year":"2018","journal-title":"Appl. Surf. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/S0003-2697(02)00257-9","article-title":"Toward optimized antibody microarrays: A comparison of current microarray support materials","volume":"309","author":"Angenendt","year":"2002","journal-title":"Anal. Biochem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1044","DOI":"10.2144\/03355rr04","article-title":"Quantitative assessment of factors affecting the sensitivity of a competitive immunomicroarray for pesticide detection","volume":"35","author":"Belleville","year":"2003","journal-title":"Biotechniques"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1016\/j.colsurfb.2008.12.037","article-title":"Enhancement of sensitivity of SPR protein microarray using a novel 3D protein immobilization","volume":"70","author":"Tanaka","year":"2009","journal-title":"Colloid Surf. B"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1002\/pmic.200390038","article-title":"Antibody microarrays: An evaluation of production parameters","volume":"3","author":"Kusnezow","year":"2003","journal-title":"Proteomics"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1016\/j.foodchem.2007.06.006","article-title":"Simultaneous detection of sulfamethazine, streptomycin, and tylosin in milk by microplate-array based SMM\u2013FIA","volume":"106","author":"Songyang","year":"2008","journal-title":"Food Chem."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Zhong, L., Zhang, W., Zer, C., Ge, K., Gao, X., and Kernstine, K.H. (2010). Protein microarray: Sensitive and effective immunodetection for drug residues. BMC Biotechnol., 10.","DOI":"10.1186\/1472-6750-10-12"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Wang, G.C., Das, C., Ledden, B., Sun, Q., Nguyen, C., and Kumar, S. (2017). Evaluation of disposable microfluidic chip design for automated and fast Immunoassays. Biomicrofluidics, 11.","DOI":"10.1063\/1.4977198"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1111\/j.1556-4029.2009.01049.x","article-title":"Development of an Antibody Hapten-Chip System for Detecting the Residues of Multiple Antibiotic Drugs","volume":"54","author":"Chen","year":"2009","journal-title":"J. Forensic Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"8925","DOI":"10.1007\/s00216-013-7311-5","article-title":"Development of a direct ELISA based on carboxy-terminal of penicillin-binding protein BlaR for the detection of beta-lactam antibiotics in foods","volume":"405","author":"Peng","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1021\/ac035028i","article-title":"Automated microarray system for the simultaneous detection of antibiotics in milk","volume":"76","author":"Knecht","year":"2004","journal-title":"Anal. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s00604-011-0548-9","article-title":"Simultaneous determination of four different antibiotic residues in honey by chemiluminescence multianalyte chip immunoassays","volume":"173","author":"Wutz","year":"2011","journal-title":"Microchim. Acta"}],"container-title":["Biosensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6374\/12\/8\/578\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:58:50Z","timestamp":1760140730000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6374\/12\/8\/578"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,29]]},"references-count":33,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["bios12080578"],"URL":"https:\/\/doi.org\/10.3390\/bios12080578","relation":{},"ISSN":["2079-6374"],"issn-type":[{"value":"2079-6374","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,29]]}}}